Cellular compartment

About: Cellular compartment is a research topic. Over the lifetime, 1082 publications have been published within this topic receiving 53794 citations. The topic is also known as: cell compartmentation.

Rapid insulin-induced exocytosis in white rat adipocytes.

Abstract: Insulin is believed to increase glucose permeability of adipocytes by regulating the incorporation of glucose transporters into the plasma membrane by exocytosis. This process involves fusion of membrane-bound cellular compartments with the plasma membrane, thus influencing the plasma membrane area. However, insulin-induced changes in plasma membrane area have not yet been demonstrated. In the present study we monitored fluorescence intensity with a confocal microscope to study the effect of insulin on adipocyte plasma membrane area. After cell isolation and adhesion to a glass cover-slip, adipocytes were stained with the dye FM1-43, a membrane area reporter. At rest, the rate of fluorescence intensity increase was initially high, but gradually stabilized at 2%/min. This steady increase in fluorescence is due to a slow rate of exocytosis coupled to endocytosis, since the removal of FM1-43 from the bath did not abolish FM1-43 fluorescence. Insulin addition caused an abrupt increase of fluorescence intensity of 4%/min, which was significantly higher than in controls. These results suggest rapid, insulin-induced incorporation of new membrane into the plasma membrane by exocytosis.

Fluctuations in cell density alter protein markers of multiple cellular compartments, confounding experimental outcomes

Abstract: The life cycle of cultured proliferating cells is characterized by fluctuations in cell population density induced by periodic subculturing. This leads to corresponding changes in micro- and macroenvironment of the cells, accompanied by altered cellular metabolism, growth rate and locomotion. Studying cell density-dependent morphological, physiological and biochemical fluctuations is relevant for understanding basic cellular mechanisms and for uncovering the intrinsic variation of commonly used tissue culture experimental models. Using multiple cell lines, we found that expression levels of the autophagic markers p62 and LC3II, and lysosomal enzyme cathepsin D were altered in highly confluent cells as a consequence of nutrient depletion and cell crowding, which led to inactivation of the mTOR signaling pathway. Furthermore, both Lamp1 and active focal adhesion kinase (FAK) were reduced in high-density cells, while chemical inhibition or deletion of FAK led to alterations in lysosomal and autophagic proteins, as well as in the mTOR signaling. This was accompanied by alterations in the Hippo signaling pathway, while cell cycle checkpoint regulator p-cdc2 remained unaffected in at least one studied cell line. On the other hand, allometric scaling of cellular compartments in growing cell populations resulted in biochemically detectable changes in the plasma membrane proteins Na+K+-ATPase and cadherin, and nuclear proteins HDAC1 and Lamin B1. Finally, we demonstrate how treatment-induced changes in cell density and corresponding modulation of susceptible proteins may lead to ambiguous experimental outcomes, or erroneous interpretation of cell culture data. Together, our data emphasize the need to recognize cell density as an important experimental variable in order to improve scientific rigor of cell culture-based studies.

The Subcellular Localization and Oligomerization Preferences of NME1/NME2 upon Radiation-Induced DNA Damage.

Abstract: Nucleoside diphosphate kinases (NDPK/NME/Nm23) are enzymes composed of subunits NME1/NDPK A and NME2/NDPK B, responsible for the maintenance of the cellular (d)NTP pool and involved in other cellular processes, such as metastasis suppression and DNA damage repair. Although eukaryotic NDPKs are active only as hexamers, it is unclear whether other NME functions require the hexameric form, and how the isoenzyme composition varies in different cellular compartments. To examine the effect of DNA damage on intracellular localization of NME1 and NME2 and the composition of NME oligomers in the nucleus and the cytoplasm, we used live-cell imaging and the FRET/FLIM technique. We showed that exogenous NME1 and NME2 proteins co-localize in the cytoplasm of non-irradiated cells, and move simultaneously to the nucleus after gamma irradiation. The FRET/FLIM experiments imply that, after DNA damage, there is a slight shift in the homomer/heteromer balance between the nucleus and the cytoplasm. Collectively, our results indicate that, after irradiation, NME1 and NME2 engage in mutual functions in the nucleus, possibly performing specific functions in their homomeric states. Finally, we demonstrated that fluorophores fused to the N-termini of NME polypeptides produce the largest FRET effect and thus recommend this orientation for use in similar studies.

Asymmetry of Plant Cell Divisions under Salt Stress

Abstract: Salt stress causes several damaging effects in plant cells. These commonly observed effects are the results of oxidative, osmotic, and toxic stresses. To ensure normal growth and development of tissues, the cellular compartments of multicellular plants have a unique system that provides the specified parameters of growth and differentiation. The cell shape and the direction of division support the steady development of the organism, the habit, and the typical shape of the organs and the whole plant. When dividing, daughter cells evenly or unevenly distribute the components of cytoplasm. Factors such as impaired osmotic regulation, exposure to toxic compounds, and imbalance in the antioxidant system cause disorders associated with the moving of organelles, distribution transformations of the endoplasmic reticulum, and the vacuolar compartment. In some cases, one can observe a different degree of plasmolysis manifestation, local changes in the density of cytoplasm. Together, these processes can cause disturbances in the direction of cell division, the formation of a phragmoplast, the formation of nuclei of daughter cells, and a violation of their fine structural organization. These processes are often accompanied by significant damage to the cytoskeleton, the formation of nonspecific structures formed by proteins of the cytoskeleton. The consequences of these processes can lead to the death of some cells or to a significant change in their morphology and properties, deformation of newly formed tissues and organs, and changes in the plant phenotype. Thus, as a result of significant violations of the cytoskeleton, causing critical destabilization of the symmetric distribution of the cell content, disturbances in the distribution of chromosomes, especially in polyploid cells, may occur, resulting in the appearance of micronuclei. Hence, the asymmetry of a certain component of the plant cell is a marker of susceptibility to abiotic damage.